Introduction: Michelia champaca L. (Magnoliaceae) is an ancient Indian medicinal plant a native of the Indian subcontinent and possessing numerous traditional uses. β-sitosterol is an important plant sterol present in Michelia which is reported to posses’ chemopreventive and adaptogenic properties. In the present study, High Performance Thin Layer Chromatography has been developed for detection, and quantification of β-sitosterol in Michelia champaca (leaves and stem-bark), after its detection and characterization initially by TLC. Methods: Increasing serial dilutions of reference standard β-sitosterol (200 to 1000 μg mL-1) were scanned at 273 nm to detect and quantify the concentrations of β-sitosterol in the test samples. Results: The estimated values obtained from the same were 791.726 μg mL-1 and 696.446 μg mL-1 for leaves and stem bark respectively. Leaves were found to be the richest source of β-sitosterol in Michelia champaca. Conclusion: The method provided a rapid and easy approach for detection and the quantitation of the bio-marker β-sitosterol. The authors also aim to validate the present method in terms of ruggedness and accuracy and undertake the isolation of β-sitosterol from the said plant.